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E timmins schiffman_physiomar12
1. The Physiological Response
of Crassostrea gigas to CO2-‐‑
Induced Ocean
Acidification
Emma Timmins-Schiffman
Carolyn Friedman
Steven Roberts
University of Washington
School of Aquatic and Fishery Sciences
Seattle, WA
3. Ocean Acidification and
the Environment
• Rising pCO2 in the atmosphere equilibrates with
ocean surface water, reducing the pH
4. Ocean Acidification and
the Environment
• Rising pCO2 in the atmosphere equilibrates with
ocean surface water, reducing the pH
• Current and projected changes in pCO2 are
unprecedented
Zeebe 2011
5. Ocean Acidification and
Bivalves
• How do changes in oceanic pCO2 and/or pH
affect the physiology of bivalves?
o C. gigas larvae show a developmental delay (Timmins-Schiffman et al., in press
Marine Biology)
o Alterations to shell growth/maintenance
o Changes in metabolic rate
o Changes in responses to other stressors
Lannig et al. 2010
15. Shell Weight
0.7
0.6
Elevated pCO2 has
Gain in Shell Weight (g)
a negative effect
0.5
on growth
0.4
0.3
0.2
0.1
0.0
400 2800 800 1200 600 1000
Treatment (µatm)
16. Proteome
• Identified 897 unique proteins across all samples
• Proteins were annotated by 453 distinct GO
categories
17. Proteome
cell cycle and
proliferation
cell organization
cell adhesion
and biogenesis
cell-‐‑cell
stress response
transport
signaling
death
protein
RNA metabolism
metabolism
signal
transduction
DNA metabolism
All proteins
developmental
processes
18. Proteome
Entire
proteome
represented
by GO terms
21. Exposure comparison
• Proteins expressed at high pCO2 are related to the
physiological processes
o Metabolism
o Oxidative stress
o Transport (proton and calcium)
o Cellular stress
o Translation
• These processes imply that the oyster is maintaining
a significant response to stress even after 1 month of
exposure
22. Exposure Comparison
15 400 µatm + MS
400 µatm
10
5
PC 2 (20.5%)
0
-5
2800 µatm + MS
-10
2800 µatm
-15
-15 -10 -5 0 5 10 15
PC 1 (49.7%)
23. Results
• MS approach expands on previous 2D gel analyses
• Shotgun sequencing allow for:
1. Identification of more proteins
2. Greater potential to detect more nuanced responses and protein
interactions
24. Conclusions
• We were able to identify hundreds of proteins in the
C. gigas gill proteome
• Even after 1 month of exposure, oysters were
maintaining an energetically costly stress response
10
8
Oyster Shell Weight (g)
6
4
400 ppm
600 ppm
2
800 ppm
1000 ppm
1200 ppm
1400 ppm
0
0 2 4 6 8 10 12
Oyster Buoyant Weight (g)
25. Further Work
• Evaluate physiological response to combined
stressors
• Comparison of transcriptomic and proteomic
responses
• Develop a more robust system for protein
identifications
26. Acknowledgements
• UWPR (Proteomics)
o Priska von Haller, Jimmy Eng, Tahmina Jahan
• UW SAFS and Biology
o Michael O’Donnell, Emily Carrington, Ken Sebens, Matt
George
o Sam White, Mackenzie Gavery, Caroline Storer, Dave Metzger
• UW Medicinal Chemistry
o Dave Goodlett, Brook Nunn
• Oyster collection and care
o Sam Garson, Ronen Elad, Joth Davis, Jason Ragan, Dustin
Johnson
• Funding
o Research: crowdfunding through RocketHub
o Travel: UW SAFS